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Fig. 6. p27Xic1 is required for muscle differentiation. Western blot for endogenous p27Xic1 protein in uninjected embryos and embryos injected with 20 ng Con Mo or 20 ng p27Xic1 Mo harvested at stage 22. Cytoskeletal β-tubulin is used as a loading control. Embryos were injected with 20 ng p27Xic1 Mo (B-D,F,H,K), 20 ng Con Mo (E,J) or 20 ng p27Xic1 Mo + 20 pg p21Cip1 (G) along with β -gal (light blue, injected side towards the left) and analyzed at stage 15 for expression of MyoD (B), Myf5 (C), MA (E) and MHC (E-G) by whole-mount in situ hybridization. Embryos injected with 20 ng p27Xic1 Mo were incubated in HUA from gastrulation until stage 22 and analyzed by whole-mount antibody staining for 12/101 expression (H,I). Embryos injected with 20 ng Con Mo (J) or p27Xic1 Mo (K) were analyzed for apoptotic cells at stage 15 by whole-mount TUNEL staining. |
